206
AKIRA MATSUNO ET AL.
Embedment of tissues fixed with 4% PFA in LR White resin
Ultrathin section mounted on Nickel grid
Hybridization with biotinylated probe at 37°C overnight
Signal detection by 20nm streptavidin gold
~
Electron microscopic observation
Fig. 2. Outline of postembedding electron microscopic in situ hybridization.
Frozen section fixed with 4% PFA
•
Hybridization with biotinylated probe at 37°C overnight
•
Signal detection by ABC-HRP-DAB
•
Osmification
•
Embedment in Epon resin
•
Ultrathin sections mounted on Nickel grids
•
Etching with 10% H202 or 4% sodium periodate
APPlicatit of primary antibody
Visualization with Protein-A colloidal gold
1
• .
. b
.
E ectron mIcroSCOPIC 0 servatlOn
Fig. 3. Outline of combined immunohistochemistry and preembedding electron microscopic in situ hybridization.
AKIRA MATSUNO ET AL.
Embedment of tissues fixed with 4% PFA in LR White resin
Ultrathin section mounted on Nickel grid
Hybridization with biotinylated probe at 37°C overnight
Signal detection by 20nm streptavidin gold
~
Electron microscopic observation
Fig. 2. Outline of postembedding electron microscopic in situ hybridization.
Frozen section fixed with 4% PFA
•
Hybridization with biotinylated probe at 37°C overnight
•
Signal detection by ABC-HRP-DAB
•
Osmification
•
Embedment in Epon resin
•
Ultrathin sections mounted on Nickel grids
•
Etching with 10% H202 or 4% sodium periodate
APPlicatit of primary antibody
Visualization with Protein-A colloidal gold
1
• .
. b
.
E ectron mIcroSCOPIC 0 servatlOn
Fig. 3. Outline of combined immunohistochemistry and preembedding electron microscopic in situ hybridization.